基于核糖体DNA ITS序列和叶绿体DNA trnL-F序列的青篱竹属Arundinaria及近缘属系统发育关系的初步研究(英文)

在青篱竹属Arundinaria 系统分类中由于对该属的形态分类标准取舍不一,至今仍存在着严重的意见分歧。本文采用PCR 扩增产物直接测序的方法分析青篱竹属Arundinaria 及近缘属(如苦竹属Pleioblastus、矢竹属Pseudosasa、少穗竹属Oligostachyum、巴山木竹属Bashania、肿节竹属Clavinodum 等)有关争议属的代表种或模式种(毛竹为外类群)等18 种竹种的核糖体DNA ITS 区段序列和叶绿体DNA trnL-F 序列。分别对ITS 序列、trnL-F 序列和ITS 与trnL-F 的组合序列进行系统发育分析。结果表明,和trnL-F 区段序列相比,核糖体DNA ITS 区段序列有较高的系统发育信息位点。通过最简约分析产生的ITS 树、trnL-F 树和ITS 与trnL-F 的组合序列树表明,所得树形基本相似。供试竹种(斑苦竹A. oleosa、仙居苦竹A. hsienchuensis、茶秆竹A. amabilis、长叶苦竹A. chino、苦竹A. amara、宜兴苦竹A. yixingensis、菲白竹A. fortunei、翠竹A. pygmaea、大明竹A. gramineus、巴山木竹A. fargesii、冷箭竹A. faberi、凤竹A. hupehense、鼓节矢竹P. japonica cv. Tsutsumiana、矢竹P. japonica、短穗竹B. densiflorum、肿节竹A. oedogonata、少穗竹A. sulcata)形成一个单系类群,且分为2 个不同的分支。图3 表2 参11。     

血皮槭叶绿体DNA非编码区差异序列筛选和分析

[目的]筛选出高变异率的叶绿体DNA序列,以进行血皮槭天然群体的谱系地理学研究。[方法]以血皮槭16个天然群体为试材,对叶绿体基因组20个非编码区序列进行测序研究,并利用筛选出的高变异率cpDNA序列初步分析了其遗传变异。[结果]序列比对和系统发育分析结果显示血皮槭cpDNA种内变异非常低,9个cpDNA序列检测到不同程度的变异位点,其中只有4个序列psbJ-petA、ndhF-rpl32、trnD-trnT和trnH-psbA表现出较高的变异水平。[结论]筛选出的4个高变异率cpDNA序列,可以在分子水平上研究血皮槭种内的系统发育、遗传变异和种群历史动态,为进一步研究濒危种血皮槭的分子谱系地理学奠定基础。     

Nutrient element contents of cutting seedlings of hybrid species （ Liriodendron chinense x tulipifera）

The cutting seedlings ofLiriodendron chinense xtulipifera were treated with the different concentrations of auxin (treatment₁: IBA of 50 g·kg⁻¹+NAA of 300 g·kg⁻¹; treatment₂; IBA of 100 g·kg⁻¹+NAA of 300 g·kg⁻¹). The biomass nutrient element contents for different organs (root, stem, leaf) of cutting seedling ofLiriodendron chinense xtulipifera were measured by the dry method, Kjeldahl method and Atomic Absorption Spectroscopy method. The result showed that the biomass of root, stem, and leaf of the cutting seedling treated with auxin was all remarkably increased. The contents of element C in root, stem and leaf had no significant difference between the control and auxin treatments, while the contents of N, P, K and Ca in stem were much lower than that in leaf and root. Variance analysis showed that for the same organ with different concentration treatment of auxin, the four nutrient elements (N, P, K, and Ca) had no significant difference in contents, while there existed significant or very significant difference in contents of the four nutrient elements in different organs with the same concentration auxin treatment. The N, P, K and Ca contents were very low in cutting seedlings; as a result, additional fertilizer should be applied to the seedlings when they were planted in the field. Foundation item: This paper was supported by Jiangsu Province Science Foundation (BE96350). Biography: ZHANG Xiao-ping, (1972-), female, Ph. Doctor in Nanjing Forestry University. Nanjing 210037, P. R. China. Responsible editor: Zhu Hong     

Construction and analysis of a subtracted cDNA library of Betula platyphylla female inflorescence

Female inflorescence of Betula platyphylla was sampled at an interval of each two days to analyze the background of gene expression in floral phase. On the basis of SMART strategy, the driver cDNA was obtained from total RNA of the last sample and the tester cDNA was from that of the others by RT-PCR which were subsequently used to construct a subtracted cDNA library. The result of the ESTs (expression sequence tags) blastX showed that the genes in the subtracted cDNA library could be mainly clustered into 5 groups related to metabolism, transportation and signal transduction, cell cycle, stress response, and regulation. The relationship between gene expression and development was also discussed.     

Identification of chloroplast DNA haplotypes ofAbies firma andA. homolepis using a polymerase chain reaction with species-specific primers

DNA sequences of the chloroplast spacer region between thetrnP andtrnW genes (234 bps) were determined for two Japanese fir species,Abies firma Sieb. et Zucc. andA. homolepis Sieb. et Zucc., using four individuals from each species. No intraspecific variation was found in either species, but interspecific sequence polymorphism was detected between the two species. The interspecific variation was three nucleotide changes, from the 148th to 150th nucleotide position. These three nucleotides were TAC inA. firma and GTA inA. homolepis, which were inverted. In order to identify the cpDNA haplotypes between the two species, this inversion polymorphism was utilized to develop a new marker. Species-specific primers were designed so that the 3′ ends of the primers would anneal to the mutation site, in order for the two haplotypes to be easily identified by a polymerase chain reaction (PCR).     

寄生小蜂对小青×黑杨树皮及杨圆蚧挥发油的行为反应

Four-armed airflow olfactometer was used to determining the foraging behavior ofPteroptrix longgiclava (Girault) (Hymenoptera: Aphelinidae) andEncarsia gigas (Tshumakova) (Hymenoptera: Aphelinidae) to the essential oils which emitted from the healthy bark ofPopulus pseudo-simonii×P. nigra, the infested bark injured byQuadraspidiotus gigas (Thiem & Gerneck), the body and scale of fixed 1st-instar-nymph ofQ. gigas. The results from these experiments showed that the volatile oils produced from the injured bark and from the scale of fixed 1st-instar-nymph had a higher attractive ability to female adults of the two species of wasps. The essential oil produced from the scale of the pest at dosages of 3–7 μL and the essential oil emitted by injured bark at dosages of 5–9 μL had a stronger alluring effect on the host searching behavior ofPteroptrix longgiclava. The essential oil from the body of fixed 1st-instar-nymph ofQ. gigas also had certain effect on the host locating effort ofPteroptrix longgiclava andEncarsia gigas. Those two wasps did not shown any reaction to the essential oil produced by the healthy bark of poplar. Foundation item: This paper was supported by National Natural Science Foundation of China (39970620) and the “TRAPOYT” Biography: CHI De-fu (1962-), male, Ph.D., Professor in Northeast Forestry University, Harbin 150040, P.R. China Responsible editor: Chai Ruihai     

New insights into the identity and origin of the causal agent of oak powdery mildew in Europe

Powdery mildew is the most common disease on oaks in Europe where it was first recorded at the beginning of the 20th century. Yet, little is known about the origin of the causal agent. In this study, we analysed the variability of the ribosomal DNA (rDNA) of the pathogen. The internal transcribed spacer (ITS) region, the 5.8S rRNA coding gene and the intergenic spacer (IGS) of the rDNA of 33 European (mostly French) samples of oak powdery mildew were amplified using polymerase chain reaction (PCR) and PCR products were subsequently sequenced. Four different haplotypes were obtained for ITS among the various samples (ITS_A, ITS_B, ITS_C and ITS_D). Each ITS sequence corresponded to a different IGS sequence. The comparison of ITS sequences obtained with sequences accessible in the GenBank database revealed very high homologies with different taxa. Of these, three taxa had already been described on oaks in Europe, i.e. Erysiphe alphitoides (100% homology with ITS_A), Erysiphe hypophylla (99.4% homology with ITS_C) and Phyllactinia guttata (97.64% homology with ITS_D). Our data also confirmed the 100% homology between ITS_A and the sequence described for Oïdium mangiferae, the agent of mango powdery mildew. The fourth haplotype, i.e. ITS_B, represented by nearly 25% samples, showed 100% homology with the recently described Erysiphe quercicola from Quercus spp. in Asia, and several tropical and sub‐tropical powdery mildew species, including Oïdium heveae, a major pathogen of rubber trees worldwide. Our results suggest that oak powdery mildew might originate from host shifts of tropical Erysiphe species introduced to Europe through infected exotic host plants.     

First report of an endophyte (Diaporthe phaseolorum var. sojae) from Kandelia candel

Mangrove endophytic fungus 1893 was isolated from Kandelia candel from an estuarine mangrove on the South China Sea Coast. Two new lactones 1893A and 1893B, together with other known compounds, have been isolated from its fermentation broth. To classify the endophyte correctly for further industrial application, a combination of morphological and molecular techniques was used to approach its identity. The endophyte was compared with similar species having trichogynes or trichogyne-like hyphae which apparently fused with antheridium-like hyphae, and perithecia initials developing from an ascogonial coil surrounded by enveloping hyphae in early developmental stages on pure culture. Further morphological characteristics on host and non-host were used for comparison with similar species when the endophyte was cultivated on leaves of Kandelia candel and Mangifera indica, respectively, which resulted in classifying the endophyte as a Phomopsis species. The ITS sequence of rDNA was used to infer its phylogenetic relationships with Phomopsis species that resembled the strain in morphology or ecology. Finally, the endophyte was identified as Diaporthe phaseolorum var. sojae based on morphological and molecular evidence. Our study is a first report of Diaporthe phaseolorum var. sojae isolated from mangrove Kandelia candel. Foundation item: The search was supported partly by the Guangzhou Natural Science Foundation (Grant No. 2007Z3-EO581), the Guangdong Provincial Natural Science Foundation (Grant No. 2007A0200300001-7), the Chinese High-Tech 863 Project (Grant No. 2006AA09Z422), and the National Natural Science Foundation of China (Grant No. 20572136). The first two authors have the same contribution.     

Development of species‐specific PCR primers on rDNA for the identification of European Armillaria species

Attempts to design species‐specific PCR primers from six European Armillaria species in the ribosomal RNA genes are reported. Primers were developed on the basis of the nucleotide sequence variability of the internal transcribed spacers (ITS) and the intergenic spacer (IGS1) of the ribosomal DNA. Four sets of primers gave specific PCR products for Armillaria tabescens, Armillaria mellea and Armillaria ostoyae. However, due to the high sequence similarities between Armillaria borealis and Armillaria ostoyae and between Armillaria cepistipes and Armillaria gallica no species specific amplification was obtained for these taxa.     

Fungal endophytes in woody roots of Douglas‐fir (Pseudotsuga menziesii) and ponderosa pine (Pinus ponderosa)

The fungal community inhabiting large woody roots of healthy conifers has not been well documented. To provide more information about such communities, a survey was conducted using increment cores from the woody roots of symptomless Douglas‐fir (Pseudotsuga menziesii) and ponderosa pine (Pinus ponderosa) growing in dry forests on the eastern slope of the Cascade Mountains in Washington state, USA. Fungal isolates were cultured on standard media, and then were identified using a combination of molecular and morphological methods. Fungal genera and species identified in this study will provide baseline data for future surveys of fungal endophytes. Examination of internal transcribed spacer (ITS1 and ITS2) and 5.8S rDNA sequences and morphology of cultured fungi identified 27 fungal genera. Two groups predominated: Byssochlamys nivea Westling (20.4% of isolations) and Umbelopsis species (10.4% of isolations). This is the first report of B. nivea within large woody roots of conifers. Both taxa have been previously identified as potential biological control agents. Although some trends were noted, this study found no significant evidence of host species or plant association effects on total recovery of fungal endophytes or recovery of specific fungal taxa.     

Population genetic diversity and regional differentiation of Chinese forest frogs (Rana chensinensis) in Heilongjiang Province

The Random Amplified Polymorphism DNA (RAPD) markers were used to study the intra-specific diversity and regional differentiation of the Chinese forest frogs (Rana chensinensis), which were sampled from the fields of 8 regions in, Heilongjiang Province. Totally 78 polymorphic DNA loci were amplified by 10 RAPD primers. By genetic distance analysis and phylogenetic tree reconstruction with the Neighbor-Joining (NJ) method the results showed that the populations of Chinese forest frogs distributed in 8 regions existed great differentiation (Average Fst=0.347, SD=0.235) while, there was the paradox between geographic distances and genetic distances. Based on geographic and geological data, a hypothesis was posed that it is very possible that the hilly lands in the downstream of the Songhua River and the Heilong River were the center of the origin of the Chinese forest frogs in Heilongjiang Province. And mainly through the Songhua River system, the Chinese forest frogs dispersed into the Songneng Plain from the Sanjiang Plain whereas the Fangzheng region became a sub-center for the western dispersion. Foundation item: This project is supported by the Natural Science Foundation of Heilongjiang Province (C9726). Biography: XIAO Xiang-hong (1960-), female, professor in College of Wildlife Resources, Northeast Forestry University, Harbin 150040, P.R. China. Responsible editor: Zhu Hong     

Possibility of grouping of Cyclobalanopsis species (Fagaceae) grown in Japan based on an analysis of several regions of chloroplast DNA

Four regions of chloroplast DNA were sequenced as a prospective genetic marker to identify Japanese representatives of Cyclobalanopsis: Quercus acuta, Q. sessilifolia, Q. salicina, Q. myrsinaefolia, Q. glauca, and Q. gilva. We found that Q. gilva was distinguished from other species based on both the trnL-trnF and trnT-trnL intergenic spacers. The evidence shows good coincidence with the fact that Q. gilva has several peculiar morphological features distinguishable from those of other species. There was no difference in trnL intron and matK. Both trnT-trnL and trnL-trnF intergenic spacers are capable of being used as genetic markers to identify Q. gilva among Cyclobalanopsis species.Part of this paper was presented at the 48th annual meeting of the Japan Wood Research Society, Shizuoka, April 3–5, 1998     

Identification of mangrove endophytic fungus 1403 (Fusarium proliferatum) based on morphological and molecular evidence

A mangrove endophytic fungus 1403 isolated from the South China Sea Coast, which is able to produce griseofulvin and anthracenediones under submerged fermentation, was compared with Fusarium genus with the similar morphological characters such as elongated, microconidium-producing conidiophores, ovoid microconidia and straight to slightly curved macroconidia. It was found that the fungus 1403 resembles pathogenic F. verticillioides (teleomophy Gibberella moniliforme) in the production of false head or chains and abundant microconidia on the aerial mycelium, but different in the occasional formation of polyphialides with relatively long as well as short monophialides, in its typical coiled hyphae and mycelia fusion. Through maximum Parsimony and Bayesian analyses, the fungus 1403 was further compared with some similar Fusarium species. The results indicated that this endophyte was identified as Fusarium proliferatum based on the analyses of partial 18S and 28S rDNA genes, ITS region, and EF-1α gene. Foundation project: This research was supported partly by the Guangzhou Natural Science Foundation (Grant No. 2007Z3-EO581), the Guangdong Provincial Natural Science Foundation (Grant No. 2007A0200300001-7; 05003268), the Chinese High-Tech 863 Project (Grant No. 2006AA09Z422), and the National Natural Science Foundation of China (Grant No. 20572136)     

Concentrations and seasonal dynamics of dissolved organic carbon in forest floors of two plantations （Castanopsis kawakamii and Cunninghamia lanceolata） in subtropical China

The concentrations and seasonal dynamics of DOC in forest floors of monoculture plantations ofCastanopsis kawakamii and Chinese fir (Cunninghamia lanceolata) were assessed in Sanming, Fujian, China (26°11′30″N, 117°26′00″E). Forest floor samples were taken in January, April, July and October in 2002 and divided into undecomposed material (horizon Oi), partially decomposed organic material (horizon Oe), and fully decomposed organic material (horizon Oa). Upon collection, DOC concentrations of samples were analyzed by a High Temperature TOC. The results showed that the annual average DOC concentration of Chinese fir (1341.7 mg·kg⁻¹) in the forest floor was higher than that ofCastanopsis kawakamii (1178.9 mg·kg⁻¹). Difference in DOC concentrations was observed among three horizons of the forest floor. DOC concentration of forest floor in the two forests was the highest in horizon Oe. Seasonal trends of DOC concentrations in different horizons of forest floors were similar and the maximal value occurred in autumn (or winter). The concentration and temporal change of DOC in studied forests were probably related to the variation in moisture, temperature, biological activity and quantity of organic matter in the forest floor. Foundation item: This study was supported by the Teaching and Research Award program for MOE P.R.C. (TRAPOYT) Biography: ZHANG Jiang-shan (1946-), male, Researcher in Fujian Normal University, Fuzhou 350007, P.R. China. Responsible editor: Zhu Hong     

Study on tracheid variation ofPicea koraiensis (nakai) in natural stand and plantation

Tracheid characteristics ofPicea koraiensis from natural stand in Liangshui area and plantation in Mao'ershan area were studied. The results of variance analysis showed that the tracheid length, diameter, and thickness of tracheid wall were significant differences between the growth rings. All those characteristics tend to increase from heart-center to bark. In natural stand, it has positive correlation between the characteristics. In plantation, the tracheid length has positive correlation with the trachied diameter, while both the trachied length and the trachied diameter has negative correlation with the thickness of trachied wall. The tracheid length and diameter have no significant difference in growth between natural stand and plantations. The thickness of tracheid wall from the plantation is a little thicker than that from the natural stand before 15 years. Foundation item: This paper was part of State 9th Five-Year Plan project of “Early Mensuration ofPicea koraiensis (nakai)”. Biography: REN Xu-qin (1973-), female, graduate student in Northeast Forestry University, Harbin 150040, P. R. China. Responsible editor: Song Funan     

Distribution, regeneration and succession ofFagus

The distribution, regeneration and succession ofFagus in China were systematically analyzed in comparison with those in Europe and in northern America. The results showed that it was very likely that China was the distribution center ofFagus, which originated in eastern Asia, migrated into northern America and then into Europe. Distribution ofFagus in China is relatively independent with seven species. The main reason for wide distribution ofFagus in Europe, but unitary species, is possibly the influence of the climate in Quaternary Period, and also related with influence of the ocean climate. This study was supported by National Natural Sciences Foundation of China (39500116 and 39670145) (Responsible Editor: Chai Ruihai)     

Bio-control trials of Chaetomium spirale ND35 against apple canker

A new endophytic antagonistic fungus, Chaetomium spirale ND35 from Populus tomentosa, was reported. The bio-control trials of C. spirale ND35 against the Valsa Canker of apple were preliminarily investigated. The results of dual culture on PDA plate showed that C. spirale ND35 was capable of strong antagonism against Valsa ceratosperma, and for inhibiting the mycelial growth of V. ceratosperma,.the crude extract of liquid culture of corn steep powder broth was more effective than that one of malt extract broth (MEB). The results of bio-control in greenhouse and field indicated that the disease incidence of apple tree treated with C. spirale ND35 was lower significantly than that treated by other methods. The re-isolation experiment suggested that C. spirale ND35 could colonize in stems and branches of apple trees successfully, and the ND35 colonization rate of the treatment with solid wheat bran culture was higher than that of corn steep powder broth, but the field experiment result the control effect of liquid culture of C. spirale ND35 was better than that of solid culture.     

Effects of elevated CO2 concentrations on photosynthesis, dark respiration and RuBPcase activity of three species seedlings in Changbai Mountain

Two-year-old seedlings ofPinus koraiensis, Pinus sylvestriformis andFraxinus mandshurica were treated in open-top chambers with elevated CO₂ concentrations (700 μL·L⁻¹, 500 μL·L⁻¹) and ambient CO₂ concentrations (350 μL·L⁻¹) in Changbai Mountain from June to Sept. in 1999 and 2001. The net photosynthetic rate, dark respiration rate, ribulose-1,5-bisphosphate carboxlase (RuBPcase) activity, and chlorophyll content were analyzed. The results indicated the RuBPcase activity of the three species seedlings increased at elevated CO₂ concentrations. The elevated CO₂ concentrations stimulated the net photosynthetic rates of three tree species exceptP. sylvestriformis grown under 500 μL·L⁻¹ CO₂ concentration. The dark respiration rates ofP. koraiensis andP. sylvestriformis increased under concentration of 700 μL·L⁻¹ CO₂, out that ofF. mandshurica decreased under both concentrations 700 μL·L⁻¹ and 500 μL·L⁻¹ CO₂. The seedlings ofF. mandshurica decreased in chlorophyll contents at elevated CO₂ concentrations. Foundation item: This paper was supported by the National Natural Science Foundation of China (No. 30070158). Knowledge Innovation Item of Chinese Academy of Sciences (KZCX2-406) and “Hundred Scientists” Project of Chinese Academy of Sciences. Biography: Zhou Yu-mei (1973-) Ph. Doctor, Assistant Research fellow Institute of Applied Ecology. Chinese Academy of Sciences. Shenyang 110016. P.R. China. Responsible editor: Song Funan     

Control of Lymantria dispar L. by biological agents

The experiment on control ofLymantria dispar L by using different kinds of biological measures, including nuclear polyhedrosis virus (NPV) ofLymantria dispar L., BtMP-342, sex-attractant as well as botanical insecticide, was carried out in the forest regions of Inner Mongolia in 2003. Two concentrations (2.632×10⁶ PIB·ml⁻¹ and 2.632×10⁷ PIB·ml⁻¹) ofLymantria dispar L. NPV were sprayed on the 2rd-instar-larvae ofL. dispar and 70% and 77.8% control effect were obtained respectively. BtMP0-342 was applied to the 3rd- and 4th-instar larvae and the control effect was around 80%. The sex-attractant provided by Canada Pacific Forestry Research Center also showed a good result in trappingL. dispar adults. The self-produced botanical insecticide, which was extracted from a kind of poisonous plant distributed in Daxing'an Mountains, China, exhibited a good control result in controlling the larvae ofL. dispar, and 82% mortality was observed when spraying primary liquid of the botanical insecticide on the 3rd–5th-instar-larvae in lab. Foundation item: This paper was supported by Natural Science Foundation of Heilongjiang Province (C0235) and Youth Foundation of Harbin City (0061218024). Biography: ZHANG Guo-cai (1964), male, Ph. Doctor Candidate, associate professor in School of Forestry, Northeast Forestry University, Harbin 150040, P. R. China. Responsible editor: Song Funan     

Nitrate reductase activity and its diurnal variation rhythm for Camptotheca acuminata seedlings

Nitrate reductase activity (NRA) in different plant organs and leaves in different positions of Camptotheca acuminata seedlings was determined by an In vivo assay, the diurnal variation rhythm of NRA in leaves of different positions was observed,and the correlations between leaf NRA, leaf area and lamina mass per unit area (LMA) were also examined. The results showed that NRA in the leaf was significantly highest, compared with that in other organs such as roots, stems and leaves. In this experiment, the 10 leaves were selected from the apex to the base of the seedlings in order. The different NRA occurred obviously in leaves of different positions of C. acuminata seedlings from the apex to the base, and NRA was higher in the 4th-6th leaves.The diurnal change rhythm of leaf NRA showed a one peak curve, and maximum NRA value appeared at about midday (at 12:30 or so). No obvious correlations between NRA and leaf area or lamina mass per unit area were observed. This study offered scientific foundation for the further research on nitrogen metabolism of C. acuminata.